Biomedical subjects

D Vanrompay

Publications and source records attributed to D Vanrompay.

At least 19 recordsLinked to original sources

PubMed2006

Animal models for the study of Chlamydia trachomatis infections in the female genital infection.

Despite intensive research on chlamydial pathogenesis and host immune responses, vaccine development has been hampered by incomplete understanding of the virulence factors and the critical factors governing protective immunity. It is unknown why certain Chlamydia trachomatis serovars cause asymptomatic or symptomatic infection or ascend to the upper genital tract. Former studies have used nonprimate monkey, mice or guinea-pig infection models. However, pigs are genetically and physiologically related to man and are susceptible to chlamydial infections. The main objective of the present study was the validation of the pig as an alternative animal model for C. trachomatis female genital tract infection.

Animals↗

PubMed2006

Description of the ICTI consortium: an integrated approach to the study of Chlamydia trachomatis infection.

The use of an integrated approach to the study of Chlamydia trachomatis infection of the female genital tract, presented at the mini-symposium "Chlamydia trachomatis infections" and described in the thesis of Joseph M. Lyons, has resulted in the creation of the ICTI consortium. The ICTI consortium is based on strong interaction and collaboration between basic scientists, clinicians, epidemiologists, and health care policy makers. This translational approach will help to further the valuable insight into the immunopathogenesis of this sexually transmitted infection (STI) and the development of new intervention strategies, including the vaccines and screening programs necessary to effectively diagnose, treat and prevent C. trachomatis infection. A background of the need for this integrated approach is presented and the goals and participants of the consortium are described.

Animals↗

PubMed2005

Pathogenic interactions between Chlamydophila psittaci and avian pneumovirus infections in turkeys.

Both Chlamydophila psittaci and avian pneumovirus (APV) are highly prevalent in Belgian turkeys and might contribute to the respiratory disease complex observed in turkeys. Initial outbreaks of chlamydiosis occur mostly at the age of 4-8 weeks, often accompanied by an APV infection in APV non-vaccinated farms. Regardless APV vaccination, breakthroughs of APV infection from 8 weeks on do occur, a period when also a second C. psittaci infection appears. Therefore, this study examined the pathogenicity of an APV superinfection in C. psittaci predisposed turkeys. Turkeys were infected with C. psittaci, APV or with C. psittaci followed by APV. Simulating the impact of an APV infection during the acute phase or latent phase of a C. psittaci infection, turkeys have been infected with APV at 1 and 5 weeks post C. psittaci infection, respectively. APV infection during the acute phase of a C. psittaci infection aggravates the severity of clinical signs, macroscopic lesions, pharyngeal APV excretion and histological tracheae lesions. In contrast, no clear interaction could be established after APV infection in latently C. psittaci infected specific pathogen-free (SPF) turkeys. This study clearly demonstrates the exacerbating role of APV during acute C. psittaci infection, which can play an important role in the respiratory disease complex of turkeys.

Acute Disease↗

PubMed2005

Protection of turkeys against Chlamydophila psittaci challenge by DNA and rMOMP vaccination and evaluation of the immunomodulating effect of 1 alpha,25-dihydroxyvitamin D(3).

Plasmid DNA expressing the major outer membrane protein (MOMP) of an avian Chlamydophila psittaci serovar D strain and recombinant MOMP (rMOMP) with or without the immunomodulating adjuvant 1 alpha,25-dihydroxyvitamin D(3) have been tested for their ability to elicit an immune response and induce protection in turkeys against challenge with the same serovar. Three vaccination strategies were compared: priming and boosting with either pcDNA1::MOMP or rMOMP and priming with pcDNA1::MOMP followed by rMOMP boosting. Turkeys primed with pcDNA1::MOMP showed significant protection against Cp. psittaci challenge, turkeys primed with rMOMP did not. The steroid hormone 1 alpha,25-dihydroxyvitamin D(3) augmented serum and mucosal antibody titres. However, higher antibody titres were not related to better protection and even had a negative effect on especially bacterial excretion.

Adjuvants, Immunologic↗

PubMed2005

Key role of Chlamydophila psittaci on Belgian turkey farms in association with other respiratory pathogens.

Two hundred turkey sera from eight Belgian and two French farms were tested for the presence of antibodies against avian pneumovirus (APV), Ornithobacterium rhinotracheale (ORT), Mycoplasma gallisepticum, Mycoplasma meleagridis and Chlamydophila psittaci. At slaughter, C. psittaci, APV and ORT antibodies were detected in 94, 34 and 6.5% of the turkeys, respectively. No antibodies against M. gallisepticum or M. meleagridis were present. Additionally, turkeys on three Belgian farms were examined from production onset until slaughter using both serology and antigen or gene detection. All farms experienced two C. psittaci infection waves, at 3-6 and 8-12 weeks of age. Each first infection wave was closely followed by an ORT infection starting at the age of 6-8 weeks, which was still detectable when the second C. psittaci infection waves started. Animals on farm A were not vaccinated against APV leading to an APV subtype B outbreak accompanying the first C. psittaci infection wave. Despite subtype A APV vaccination on farms B and C, the second C. psittaci infection waves were accompanied (farm B) or followed (farm C) by a subtype B APV infection. On all farms respiratory signs always appeared together with a proven C. psittaci, APV and/or ORT infection. This study suggests an association between C. psittaci, APV and ORT, and indicates the multi-factorial aetiology of respiratory infections in commercial turkeys. All three pathogens should be considered when developing prevention strategies for respiratory disease.

Age Factors↗

PubMed2004

Priming of piglets against enterotoxigenic E. coli F4 fimbriae by immunisation with FAEG DNA.

Early vaccination is necessary to protect pigs against postweaning diarrhoea caused by enterotoxigenic Escherichia coli (ETEC). However, at present no commercial vaccine allows successful vaccination. This is partly due to the presence of maternally derived antibodies. Since DNA vaccines are suggested to be superior to protein vaccines in young animals with maternal antibodies, we determined whether the fimbrial adhesin (FaeG) of F4ac(+) ETEC could be used as a plasmid DNA vaccine to prime piglets in a heterologous prime-boost approach. Hereto, pcDNA1/faeG19 was constructed and expression of rFaeG in Cos-7 cells was demonstrated. Thereafter, pigs were immunised (days 0, 21 and 42) intramuscularly by injection or intradermally by gene gun and humoral and cellular immune responses were analysed. Even though responses were low, results demonstrated that intramuscular injection was superior to gene gun delivery for priming the humoral immune response since higher antibody titres were raised, whereas gene gun delivery better induced a cellular response, evaluated by a lymphocyte proliferation assay. Effective priming of the humoral immune response was evidenced by high IgG titres 1 week after a protein boost with purified F4. The low responses to the pcDNA1/faeG19 DNA vaccination suggest that delivery of the DNA and/or the expression of the faeG gene should be improved.

Animals↗

PubMed2004

Influence of maternal antibodies on Chlamydophila psittaci-specific immune responses in turkeys elicited by naked DNA.

Plasmid DNA (pcDNA1::MOMP D) expressing the major outer membrane protein (MOMP) of an avian Chlamydophila psittaci serovar D strain was tested for its ability to induce protective immunity against C. psittaci challenge in the presence of maternal antibodies. A combined parenteral (intramuscular injection) and mucosal route (DNA drops administered to the nares) of DNA inoculation was used. Following pcDNA1::MOMP vaccination, both T helper and B cell memory were primed. However, high maternal antibodies titres affected the induction of vaccine-specific antibody responses as assessed by MOMP-specific antibody levels in enzyme-linked immunosorbent assay (ELISA). Cell-mediated immunity was unaltered as demonstrated by the significantly heightened proliferative responses of peripheral blood lymphocytes (PBL) following vaccination. DNA vaccination could significantly reduce clinical symptoms, pharyngeal and cloacal excretion as well as Chlamydophila replication, even in the presence of maternal antibodies.

Administration, Intranasal↗

PubMed2003

Missing links in the divergence of Chlamydophila abortus from Chlamydophila psittaci.

Pathological and serological evidence and DNA-DNA reassociation data indicate that Chlamydophila psittaci and Chlamydophila abortus are separate species. C. psittaci causes avian systemic disease and C. abortus causes abortion. Both previously belonged to Chlamydia psittaci are associated with zoonotic and enzootic outbreaks. Genetic studies suggest that they are closely related and because of the recent availability of diverse C. psittaci strains and comparative data for several genes, it was possible to explore this relationship. The parrot C. psittaci strain 84/2334 was found to have DNA sequences that were identical to an extrachromosomal plasmid in duck C. psittaci strain N352, to rnpB in strain R54 from a brown skua and to the rrn intergenic spacer in parakeet strain Prk/Daruma (from Germany, Antarctica and Japan, respectively). Analysis of ompA and the rrn spacer revealed progressive diversification of the strains, with 84/2334 resembling what might have been a recent ancestor of C. abortus. Another C. psittaci strain (VS225) showed evidence of having undergone convergent evolution towards the C. abortus-like genotype, whereas strain R54 diverged independently. For the first time, these studies link C. abortus in an evolutionary context to the C. psittaci lineage. It has been concluded that C. abortus diverged from C. psittaci, and so strain R54 was designated a C. psittaci strain. It is recommended that characterization of C. psittaci and C. abortus strains should utilize more than a single method and more than a single gene.

Abortion, Veterinary↗

PubMed2001

Comparative analysis of porcine cytokine production by mRNA and protein detection.

To analyze the correlation between cytokine mRNA transcription and secretion, interleukin-10 (IL-10) and interferon-gamma (IFN-gamma) were quantified by enzyme-linked immunosorbent assay and IL-2 by bioassay and compared with their mRNA levels, determined by reverse transcription polymerase chain reaction (RT-PCR). For this purpose, peripheral blood monomorphonuclear cells (PBMC) were stimulated in vitro with the lectins pokeweed mitogen (PWM), phytohemagglutinin (PHA) and concanavalin A (ConA), respectively, and with F4 fimbriae in an antigen-specific assay. Analyses were performed 4, 8, 16, 24 and 48 h after stimulation on the stimulated PBMC for mRNA and on the respective culture supernatants for proteins. RT-PCR products were quantified by densitometric scanning of the electrophoresis bands and related to the band intensity of the housekeeping gene, cyclophilin. Low levels of IL-2, IL-4 and IFN-gamma mRNA were detected in unstimulated PBMC. Stimulation with all three mitogens (PWM, ConA, PHA) led to an increase in mRNA transcription. In contrast, substantial IL-10 mRNA levels were detected in both unstimulated and stimulated cells with practically no difference between the three mitogens used. IL-2 mRNA expression tended to peak after 8-16 h for all three mitogens. The cells stimulated with PWM and ConA showed higher levels of gene expression for IFN-gamma and lower for IL-4 then the cells stimulated with PHA, however, differences were not statistically significant. For cells stimulated with F4 fimbriae only the IFN-gamma mRNA expression increased with an early peak at 8h post-stimulation. The analysis of the culture supernatants for secreted cytokines revealed a correlation between the levels of mRNA transcription and the respective secreted cytokines during the first 24h of stimulation. After 24h of stimulation, however, a decrease in IFN-gamma and IL-2 mRNA levels was accompanied by an increase or a less pronounced decrease in cytokine concentration; only the ConA induced IL-2 mRNA and protein concentration slopes showed similar profiles. In conclusion, similar cytokine production profiles as defined by mRNA and protein, respectively are obtained only during the first 24h after stimulation of the cell cultures.

Animals↗

PubMed2001

Protection of turkeys against Chlamydophila psittaci challenge by parenteral and mucosal inoculations and the effect of turkey interferon-gamma on genetic immunization.

Plasmid DNA (pcDNA1::MOMP A) expressing the major outer membrane protein of an avian Chlamydophila psittaci serovar A strain was tested for its ability to induce protective immunity against challenge with the same C. psittaci serovar. A combined parenteral (intramuscular injection) and mucosal route (DNA drops administered to the nares) of DNA inoculation was compared to three other, different routes of administration (intramuscular inoculation, DNA drops administered to the nares and aerosol immunization). In addition, the effect of turkey interferon gamma (tIFN-gamma) on intramuscular immunization was evaluated by co-expressing pCIneo::tIFN-gamma. A significant level of protection was observed in turkeys immunized via the combined parenteral/mucosal route, the intramuscular route or by aerosol. Severe clinical signs and lesions were observed in the non-vaccinated control groups, in 80% of turkeys inoculated with a mixture of pcDNA1::MOMP A and pCIneo::tIFN-gamma, and in 60% of turkeys vaccinated with DNA drops administered to the nares. The use of MOMP-based DNA vaccination as a means of preventing severe clinical signs and lesions in a turkey model of C. psittaci infection was demonstrated, as was down-regulation of the immune response by co-expression of tIFN-gamma.

Animals↗

PubMed2001

Genetic immunization for Chlamydia psittaci.

Plasmid DNA expressing the major outer membrane protein (MOMP) of an avian Chlamydia psittaci serovar A strain, has been tested for its ability to raise an immune response and induce protection against challenge with the same serovar. A combined parenteral (intramuscular injection) and mucosal route (DNA drops administered to the nares) of DNA inoculation was compared to gene gun-based immunization. The gene gun delivery of pcDNA1/MOMP as well as the intramuscular-intranasal DNA delivery primed both T-helper and B-cell memory although rMOMP-expressing cells did not induce high antibody responses. Evidence for the priming of the memory was provided by the fact that the pcDNA1/MOMP inoculations raised antibodies belonging to the IgG and not IgM isotype. However, in response to challenge only 5 out of 15 vaccinated turkeys showed four fold increases in serum IgG after challenge. By contrast, evidence for the priming of T-cell memory in response to challenge was found in all vaccinated turkeys as shown by the significantly heightened proliferative responses of peripheral blood lymphocytes following vaccination. Both immunization methods produced similar serological and lymphocyte proliferative responses. Notwithstanding the immunization method, a significant level of protection was observed in all pcDNA1/MOMP immunized turkeys. The efficacy of MOMP-based DNA vaccination as a means of preventing severe clinical signs, lesions and chlamydia excretion in a turkey model of Chlamydia psittaci infection was demonstrated.

Animals↗

PubMed2000

Avian chlamydiosis.

Avian chlamydiosis (AC) can be economically devastating to producers and a serious public health problem. Most infections in humans are due to exposure to psittacine birds and pigeons; however, outbreaks resulting in severe disease and even death do occur in abattoir workers following processing of infected flocks. The disease occurs primarily in turkeys and ducks, but can affect all types of poultry. In poultry, the disease varies from one producing high morbidity and mortality to one that is asymptomatic. Farm workers and abattoir workers are at risk following exposure to either extreme. Although outbreaks of AC have declined since the 1970s, some parts of the world are now experiencing a rise in incidence. Whether the initial decrease was due to changes in production methods or to the increased use of antibiotics is not known. The mechanism for introduction of the disease into a flock or area is poorly understood. Wild birds are often infected by the same strains as domestic flocks and are therefore thought to play a major role in introduction. Data also indicate that vertical transmission may occur. Persistently infected carrier birds are known to be a source of chlamydiosis in the pet bird industry, but have not been confirmed as a source of infection in poultry flocks.

Animals↗

PubMed1999

Protection of turkeys against Chlamydia psittaci challenge by gene gun-based DNA immunizations.

Particle-mediated (Helios Gene Gun) transfer to the turkey epidermis of plasmid DNA expressing the major outer membrane protein (MOMP) of an avian Chlamydia psittaci strain was evaluated for its ability to raise an immune response and protection against challenge with the homologous strain. In turkeys, the delivery of pcDNA1/MOMP coated onto 0.6 microm gold beads was the most efficient compared to immunisations using 1.0 or 1.6 microm gold beads. The delivery of as little as 1 microg pcDNA1/MOMP coated onto 0.6 microm gold beads was efficient. Immunisation with 1.0 microm gold beads required twice more (2 microg) DNA to achieve comparable results. The use of 2 microg DNA coated onto 1.6 microm gold beads had no effects. The gene gun delivery both primed T-helper and B-cell memory although recombinant MOMP-expressing cells did not induce high-titre antibody responses. The significance of gene gun-based DNA immunisation as a means of preventing severe clinical signs, lesions and chlamydia excretion in a turkey model of Chlamydia psittaci infection was demonstrated.

Animals↗

PubMed1999

Turkeys are protected from infection with Chlamydia psittaci by plasmid DNA vaccination against the major outer membrane protein.

Plasmid DNA expressing the major outer membrane protein (MOMP) of an avian Chlamydia psittaci serovar A strain has been tested for its ability to raise an immune response and induce protection against challenge with the same serovar. A combined parenteral (intramuscular injection) and mucosal route (DNA drops administered to the nares) of DNA inoculation was compared with gene gun-based immunization. The gene gun delivery of pcDNA1/MOMP as well as the intramuscular-intranasal DNA delivery primed both T-helper and B cell memory, although rMOMP-expressing cells did not induce high antibody responses. Evidence for the priming of the memory was provided by the fact that the pcDNA1/MOMP inoculations raised antibodies belonging to the IgG and not IgM isotype. However, in response to challenge only five out of 15 vaccinated turkeys showed four-fold increases in serum IgG after challenge. By contrast, evidence for the priming of T cell memory in response to challenge was found in all vaccinated turkeys, as shown by the significantly heightened proliferative responses of peripheral blood lymphocytes following vaccination. Both immunization methods produced similar serological and lymphocyte proliferative responses. Notwithstanding the immunization method, a significant level of protection was observed in all pcDNA1/MOMP-immunized turkeys. The efficacy of MOMP-based DNA vaccination as a means of preventing severe clinical signs, lesions and chlamydia excretion in a turkey model of C. psittaci infection was demonstrated.

Animals↗

PubMed1998

High-level expression of Chlamydia psittaci major outer membrane protein in COS cells and in skeletal muscles of turkeys.

The omp1 genes encoding the major outer membrane proteins (MOMPs) of avian Chlamydia psittaci serovar A and D strains were cloned and sequenced. The nucleotide sequences of the avian C. psittaci serovar A and D MOMP genes were found to be 98.9 and 87.8% identical, respectively, to that of the avian C. psittaci serovar A strain 6BC, 84.6 and 99.8% identical to that of the avian C. psittaci serovar D strain NJ1, 79.1 and 81.1% identical to that of the C. psittaci guinea pig inclusion conjunctivitis strain, 60.9 and 62.5% identical to that of the Chlamydia trachomatis L2 strain, and 57.5 and 60.4% identical to that of the Chlamydia pneumoniae IOL-207 strain. The serovar A or D MOMPs were cloned in the mammalian expression plasmid pcDNA1. When pcDNA1/MOMP A or pcDNA1/MOMP D was introduced into COS7 cells, a 40-kDa protein that was identical in size, antigenicity, and electrophoretic mobility to native MOMP was produced. Recombinant MOMP (rMOMP) was located in the cytoplasm of transfected COS7 cells as well as in the plasma membrane and was immunoaccessible. Intramuscular administration of pcDNA1/MOMP in specific-pathogen-free turkeys resulted in local expression of rMOMP in its native conformation, after which anti-MOMP antibodies appeared in the serum.

Amino Acid Sequence↗

PubMed1997

The prevalence of Chlamydia psittaci infections in Belgian commercial turkey poults.

The prevalence of Chlamydia psittaci infections in Belgian commercial turkey poults was examined and a follow-up study of one Belgian turkey flock was performed. Sera were examined for the presence of anti-chlamydia antibodies by immunoblotting. Cloacal and conjunctival swab smears and lung impression smears were examined for the presence of chlamydial antigen using the IMAGEN Chlamydia immunofluorescence test. Anti-chlamydia antibodies were found in 90 of 100 sera collected at slaughter from turkeys raised during the summer of 1992. The following winter, 73 of 100 sera reacted positively. On all twenty farms examined during 1992, turkeys were positive for anti-chlamydial antibodies. During 1993, chlamydial antigen was detected in swabs from 20 of 40 slaughterhouse turkeys tested. Antigen was found more often in the cloaca than in the conjunctiva. Chlamydial antigen was detected in samples from each of the 4 farms examined. The follow-up study on a turkey farm, sampling the birds at weekly intervals from one week old until 12 weeks of age, revealed that chlamydial antigen and anti-chlamydial antibodies were present during the whole period. During 1994, chlamydial antigen was detected in 45 of 60 lungs from slaughterhouse turkeys from all of 6 farms. During 1995, chlamydial antigen was detected in 41 of 54 lungs of 6 week old commercial turkey poults. The results of the present study indicate that Chlamydia psittaci infections are highly prevalent amongst Belgian commercial turkey poults with apparently little seasonal or year-to-year variation and that turkeys can contract the infection at an early age.

Animals↗

PubMed1997

Characterization of avian Chlamydia psittaci strains using omp1 restriction mapping and serovar-specific monoclonal antibodies.

In the present study, 60 avian Chlamydia psittaci isolates were characterized using restriction fragment length polymorphism as well as serovar-specific monoclonal antibodies, enabling a comparison between the two characterization methods. Sixty avian C. psittaci isolates were characterized by Alul restriction mapping of the major outer membrane protein gene omp1 obtained after amplification by the polymerase chain reaction. The 60 avian C. psittaci strains were also characterized using serovar-specific monoclonal antibodies in a microimmunofluorescence test. Digestion of 60 avian C. psittaci omp1 amplicons by Alul generated 5 of the 6 known distinct restriction patterns (A, B, D, E and F). Restriction pattern C was not observed. Serotyping revealed 4 avian C. psittaci serovars (A, B, C and D). None of the 60 isolates was typed as serovar E. AluI restriction patterns A, B, D and E corresponded in 98% of the cases to serovars A, B, C and D, respectively. One isolate, classified as serovar A, generated restriction pattern F instead of A. Genotyping enabled a more precise differentiation of avian C. psittaci serovar A strains. Serovar A strains were divided into two groups according to their Alul restriction pattern (A or F). For epidemiological studies, genotyping can thus be a highly valuable alternative to serotyping, especially when applied directly to the clinical samples.

Animals↗

PubMed1996

Ultrastructural changes in avian Chlamydia psittaci serovar A-, B-, and D-infected Buffalo Green Monkey cells.

In order to find an explanation for the observed differences in levels of pathogenicity in turkeys of Chlamydia psittaci 84/55 (avian serovar A), 89/1326 (avian serovar B), 92/1293 (avian serovar D), and the Texas Turkey strain (avian serovar D) (P.B. Wyrick, J. Choong, S.T. Knight, D. Goyeau, E.S. Stuart, and A.B. MacDonald, Immunol. Infect. Dis. 4:131-141, 1994), the reproductive cycles of organisms of the four strains were studied in Buffalo Green Monkey cells by transmission electron microscopy, immunoelectron microscopy, and flow cytometry. Organisms of strains most pathogenic in turkeys, namely, the serovar A strain and the 92/1293 serovar D strain, (i) replicated faster, since at 50 h postinoculation significantly larger inclusions with more numerous infectious organisms were observed than with the less pathogenic strains; (ii) were often found devoid of inclusion membranes scattered throughout the cytoplasms; and (iii) induced severe degenerative changes in Buffalo Green Monkey cells. By immunoelectron microscopy and flow cytometry, chlamydial antigens could not be detected in the plasma membranes of infected host cells. However, the presence of chlamydial antigens in inclusion membranes was demonstrated by immunoelectron microscopy.

Animals↗